Summary: Activation of human monocytes by bacterial endotoxin, lipopolysaccharide (LPS), induces expression of many cytokines, including tumor necrosis factor (TNF), interleukin-1 (IL-1), IL-6 and IL-10. IL-10 expression is delayed relative to that of TNF, IL-1 and IL-6. Furthermore, IL-10 feedback inhibits expression of TNF, IL-1 and IL-6, thus providing an efficient autocrine mechanism for controlling proinflammatory cytokine production in monocytes. In this project, we are examining the mechanism by which IL-10 down-regulates production of cytokines such as TNF and IL-1 in endotoxin-stimulated monocytes. We are also evaluating the effects of IL-10 on signal transduction events that are activated by cytokines such as IFN-gamma (IFN-g) and IL-4. We have found that IL-10 inhibits activation and gene expression induced by IL-4 and IFN-gamma. We have also determined that the ability of IL-10 to inhibit IL-4-inducible gene expression is a consequence of decreased tyrosine phosphorylation and nuclear translocation of the IL-4-inducible transcription factor, STAT6. We are now examining the role of a novel family of JAK/STAT inhibitory genes, the Suppressors of Cytokine Signaling (SOCS) genes, in mediating these IL-10-inducible inhibitory effects. We have found that IL-10 selectively induces expression of one member of this newly identified gene family, SOCS-3. Forced expression of SOCS-3 in a macrophage cell line markedly inhibits induction of STAT activity by several cytokines, including IFN-gamma, GM-CSF and IL-4. Therefore, the ability of IL-10 to antagonize cytokine-inducible gene expression appears to be associated with its ability to induce rapid expression of the SOCS-3 gene. In related studies, we are also examining the biological activities of several newly identified IL-10 homologues. One of these, IL-19, shares approximately 21% amino acid homology with IL-10, and may utilize at least one of the two receptor chains that form the functional IL-10 receptor complex. We have found that expression of the IL-19 gene is upregulated by many of the same agents that induce IL-10 gene expression in monocytes. However, unlike the IL-10 gene, IL-19 is not expressed in activated T cells. Another IL-10 homologue, IL-TIF (IL-10-related T cell-derived inducible factor), has also recently been described. Together with scientists at the University of Medicine & Dentistry of New Jersey (UMDNJ), we have identified and characterized the gene encoding the ligand-binding chain of the receptor for this novel IL-10-related cytokine, IL-TIF. This receptor chain heterodimerizes with the IL-10R-beta chain (IL-10R2) to form a functional IL-TIF receptor complex. Therefore, IL-10R2 is component of both the IL-10 and IL-TIF receptors. We have recently found that the IL-TIF receptor is expressed at high levels in liver and kidney, but not in hematopoietic tissues such as the spleen and thymus. This suggests a possible functional role for this receptor in regulating gene expression in these tissues.